Confluence Data

BT474 human breast carcinoma cells were obtained from American Type Culture Collection (ATCC, Manassas, VA). The cells were grown in Dulbecco's modified eagle medium (DMEM, Thermo Fisher Scientific Inc., Waltham, MA) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO) 1% L-glutamine (Thermo Fisher Scientific Inc.), and 1% Penicillin Streptomycin (Thermo Fisher Scientific Inc.) in 5% CO_2 and air at 37C. <br>D-(+)-glucose solution (Sigma-Aldrich) was added to glucose free DMEM to yield media with glucose concentrations of 2, 5, and 10 mM, which was not replenished during the experiments. For each glucose level there were three initial seeding densities, each with four replicates. Cells were seeded at a density of 3.5x10^4, 5.0x10^4 and 6.0x10^4 cells/well on a 96-well tissue culture plate.